Estrogen stimulates dimethylarginine dimethylaminohydrolase activity and the metabolism of asymmetric dimethylarginine.

Background— Experimental evidence suggests that estrogens stimulate the production of nitric oxide (NO) by vascular endothelial cells. This effect has been attributed to increased expression and enzymatic activity of both the constitutive and inducible isoforms of NO synthase. In this study, we have investigated whether estrogens regulate the metabolism or release of asymmetric dimethylarginine (ADMA), an endogenous inhibitor of NO synthase. Methods and Results— The concentration of ADMA in the plasma of 15 postmenopausal women was 0.722±0.04 μmol/L (mean±SEM). Two weeks after subcutaneous implantation with estradiol, there was an increase in plasma estradiol concentration from 0.693±0.075 to 0.81±87 nmol/L, which was accompanied by a significant fall in plasma ADMA concentration to 0.588±0.03 μmol/L (P=0.006). Human and murine endothelial cell lines previously cultured in estrogen-free medium and then exposed to 17β-estradiol showed a dose-dependent decrease in the release of ADMA. This reached statistical significance at 10−14 mol/L 17β-estradiol and was accompanied by a corresponding increase in the activity of dimethylarginine dimethylaminohydrolase (DDAH), an enzyme that catalyzes the metabolism of ADMA. Conclusions— We have demonstrated that estrogens can alter the catabolism and release of ADMA in vitro and reduce the circulating concentration in vivo. We therefore propose that increased DDAH activity and the subsequent fall in ADMA could contribute to the positive effect of estrogen on NO synthesis

Influence of elevated carbon dioxide concentrations on methane emission and its associated soil microflora in rice ecosystem

The dynamics of methane emission and its associated soil microflora in rice ecosystem as a response to elevated CO2 concentrations were studied in open top chamber (OTC) conditions. The treatments consisted of three levels of CO2 (396, 550 and 750 µmol mol-1) and three levels of nitrogen (0, 150 and 200 kg ha-1) and replicated five times in a completely randomized design. The data showed that elevated [CO2] significantly (P ? 0.01) increased the DOC throughout the cropping period with the values ranging from 533 to 722 mg L-1 and 368 to 501 mg L-1 in C750 and Camb, respectively. Methane emission rates were monitored regularly during the experiment period and it was revealed that elevated [CO2] had increased the methane emissions regardless of stages of crop growth.  It was observed that methane emissions were significantly higher under [CO2] of 750 µmol mol-1 by 33 to 54 per cent over the ambient [CO2] of 396 µmol mol-1. Consistent with the observed increases in methane flux, the enumeration of methanogens showed a significant (P ? 0.01) increase under elevated [CO2] with the population ranging from 5.7 to 20.1 x 104 CFU g-1 of dry soil and 5.1 to 16.9 x 104 CFU g-1 of dry soil under C750 and Camb concentrations, respectively. Interestingly, even though higher methanotrophs population was recorded under elevated [CO2], it could not circumvent the methane emission. Overall, the results of OTC studies suggest that methane mitigation strategies need to be explored for the future high CO2 environments.

Packed Red Blood Cells Are an Abundant and Proximate Potential Source of Nitric Oxide Synthase Inhibition

<div><p>Objective</p><p>We determined, for packed red blood cells (PRBC) and fresh frozen plasma, the maximum content, and ability to release the endogenous nitric oxide synthase (NOS) inhibitors asymmetric dimethylarginine (ADMA) and monomethylarginine (LNMMA).</p><p>Background</p><p>ADMA and LNMMA are near equipotent NOS inhibitors forming blood’s total NOS inhibitory content. The balance between removal from, and addition to plasma determines their free concentrations. Removal from plasma is by well-characterized specific hydrolases while formation is restricted to posttranslational protein methylation. When released into plasma they can readily enter endothelial cells and inhibit NOS. Fresh rat and human whole blood contain substantial protein incorporated ADMA however; the maximum content of ADMA and LNMMA in PRBC and fresh frozen plasma has not been determined.</p><p>Methods</p><p>We measured total (free and protein incorporated) ADMA and LNMMA content in PRBCs and fresh frozen plasma, as well as their incubation induced release, using HPLC with fluorescence detection. We tested the hypothesis that PRBC and fresh frozen plasma contain substantial inhibitory methylarginines that can be released chemically by complete <i>in vitro</i> acid hydrolysis or physiologically at 37°C by enzymatic blood proteolysis.</p><p>Results</p><p><i>In vitro</i> strong-acid-hydrolysis revealed a large PRBC reservoir of ADMA (54.5 ± 9.7 µM) and LNMMA (58.9 ± 28.9 μM) that persisted over 42-d at 6° or -80°C. <i>In vitro</i> 5h incubation at 37°C nearly doubled free ADMA and LNMMNA concentration from PRBCs while no change was detected in fresh frozen plasma.</p><p>Conclusion</p><p>The compelling physiological ramifications are that regardless of storage age, 1) PRBCs can rapidly release pathologically relevant quantities of ADMA and LNMMA when incubated and 2) PRBCs have a protein-incorporated inhibitory methylarginines reservoir 100 times that of normal free inhibitory methylarginines in blood and thus could represent a clinically relevant and proximate risk for iatrogenic NOS inhibition upon transfusion.</p></div

Efecto de la temperatura sobre las propiedades de los materiales carbonosos provenientes de la pirólisis de lodos residuales y su aplicación como sorbentes de iones en soluciones bimetalicas de cadmio y cromo

Como resultados de los análisis de los materiales carbonosos y en base lo encontrado en la literatura (propiedades) se eligieron tres materiales carbonosos (MC 500, MC600 y MC 700) para evaluar la capacidad de sorción con una solución bimetálica de cadmio y cromo. Primero se realizo la cinética de sorción para determinar el tiempo de equilibrio y el porcentaje remoción de cada uno de los metales. Posteriormente, los datos experimentales obtenidos los tres materiales carbonosos se ajustaron a los diferentes modelos cinéticos linealizados (primer orden, segundo orden y pseudo- ecuación de segundo orden) para conocer el tipo de adsorción que se lleva acabo,En este trabajo de investigación se obtuvieron 5 materiales carbonosos provenientes de la pirólisis de lodos residuales a diferentes temperaturas 400, 500, 600, 700 y 750 ºC basados en el análisis termogravimétrico (TGA) y térmico diferencial (DTA) del lodo residual seco, el incremento en la temperatura de pirólisis le confirió al material carbonoso diversas propiedades estructurales, morfológicas y texturales. Estas variaciones en las propiedades se determinaron a partir de las siguientes técnicas: Micronanalisis elemental por Dispersión de Energía de Rayos-X (EDS), Espectroscopia Infrarroja (IR), Microscopia Electrónica de Barrido (MEB) y Fisisorción de Nitrógeno.UAE

Comparison of analytical methodologies to diagnose soils with basic amendments in the argentine temperate area

Se desconoce si los protocolos de pH y capacidad de intercambio catiónico usados actualmente son igualmente apropiados para el seguimiento de la evolución de la acidez del suelo con encalado previo. Los objetivos del trabajo fueron analizar la capacidad diagnóstica a través del tiempo del pH actual y potencial en 2 diluciones y 2 protocolos para la determinación de la capacidad de intercambio catiónico y las bases intercambiables sobre 2 suelos ácidos de la provincia de Buenos Aires, tratados con diferentes tipos y dosis de enmiendas básicas. Las unidades experimentales fueron macetas de 750 g mantenidas a 80- 90% de su capacidad de campo; el tiempo de incubación fue 3-6-9-12 meses. Los correctores fueron: aplicación equivalente a 0, 750, 1.500 y 2.000 kg ha-1 de CaCO3 y de CaCO3-MgCO3, respectivamente. Se determinó potenciométricamente pH actual y potencial en relación suelo:agua y KCl, respectivamente en 1:2,5, y 1:1; capacidad de intercambio catiónico (CIC) y cationes intercambiables mediante acetato de NH4 1M pH 7 como extractante y CIC no bufferada (CICnb) mediante NH4Cl 0,2 M al pH del suelo. Los 4 protocolos utilizados para evaluar el pH están altamente asociados entre sí. Todos ellos fueron capaces de discriminar las dosis de encalado, aunque el pH actual 1:2,5 fue el que presentó las mayores diferencias. Sin embargo, las medidas de pH actual son más fluctuantes que las de pH potencial, a cualquier dilución. Los resultados señalan que una anticipación de 6 meses en la práctica del encalado podría ser suficiente en cultivos sensibles en suelos de similar pH a los estudiados. Ninguno de los métodos de determinación de la capacidad de intercambio catiónico, ni la saturación de bases permitió registrar un incremento de la medida ante el encalado, ni discriminar el efecto de dosis.It is unknown whether pH and cation exchange capacity protocols currently used are also suitable for monitoring limed soil acidity evolution. The objectives of this study were to analyze the diagnostic performance over time of current and potential pH tested in two acid soils of the Buenos Aires province, treated with different doses of basic amendments, using two dilutions and two cation exchange capacity and exchangeable bases protocols. The experimental units were 750 g pots maintained at 80-90% of soil field capacity; the incubation time was of 3, 6, 9, and 12 months. The corrector applications were equivalent to 0,750, 1.500 and 2.000 kg ha-1 of CaCO3 and CaCO3-MgCO3 respectively. Current and potencial pH were determined potentiometrically in a 1:2,5 water and 1:1 KCl relation respectively. Also cationic exchange capacity (CEC) with 1M NH4 acetate pH 7 as extractant and cationic exchange capacity not buffered (CICnb) with 0.2 M NH4Cl extractant at soil´s pH were determined. The four protocols used to evaluate the pH were highly associated. The four pH measures were able to discriminate liming rates although the current pH 1:2,5 presented the greatest differences. However, actual pH measurements were more volatile than the potential pH, at any dilution. The results indicate that 6-month anticipation liming may be sufficient for sensitive crops in soils with similar pH to those studied here. None of the determination methods for the cation exchange capacity and the saturation of the bases resulted in a size increase before liming or to discriminate the effect of each rate.Facultad de Ciencias Agrarias y Forestale

Comparison of analytical methodologies to diagnose soils with basic amendments in the argentine temperate area

Se desconoce si los protocolos de pH y capacidad de intercambio catiónico usados actualmente son igualmente apropiados para el seguimiento de la evolución de la acidez del suelo con encalado previo. Los objetivos del trabajo fueron analizar la capacidad diagnóstica a través del tiempo del pH actual y potencial en 2 diluciones y 2 protocolos para la determinación de la capacidad de intercambio catiónico y las bases intercambiables sobre 2 suelos ácidos de la provincia de Buenos Aires, tratados con diferentes tipos y dosis de enmiendas básicas. Las unidades experimentales fueron macetas de 750 g mantenidas a 80- 90% de su capacidad de campo; el tiempo de incubación fue 3-6-9-12 meses. Los correctores fueron: aplicación equivalente a 0, 750, 1.500 y 2.000 kg ha-1 de CaCO3 y de CaCO3-MgCO3, respectivamente. Se determinó potenciométricamente pH actual y potencial en relación suelo:agua y KCl, respectivamente en 1:2,5, y 1:1; capacidad de intercambio catiónico (CIC) y cationes intercambiables mediante acetato de NH4 1M pH 7 como extractante y CIC no bufferada (CICnb) mediante NH4Cl 0,2 M al pH del suelo. Los 4 protocolos utilizados para evaluar el pH están altamente asociados entre sí. Todos ellos fueron capaces de discriminar las dosis de encalado, aunque el pH actual 1:2,5 fue el que presentó las mayores diferencias. Sin embargo, las medidas de pH actual son más fluctuantes que las de pH potencial, a cualquier dilución. Los resultados señalan que una anticipación de 6 meses en la práctica del encalado podría ser suficiente en cultivos sensibles en suelos de similar pH a los estudiados. Ninguno de los métodos de determinación de la capacidad de intercambio catiónico, ni la saturación de bases permitió registrar un incremento de la medida ante el encalado, ni discriminar el efecto de dosis.It is unknown whether pH and cation exchange capacity protocols currently used are also suitable for monitoring limed soil acidity evolution. The objectives of this study were to analyze the diagnostic performance over time of current and potential pH tested in two acid soils of the Buenos Aires province, treated with different doses of basic amendments, using two dilutions and two cation exchange capacity and exchangeable bases protocols. The experimental units were 750 g pots maintained at 80-90% of soil field capacity; the incubation time was of 3, 6, 9, and 12 months. The corrector applications were equivalent to 0,750, 1.500 and 2.000 kg ha-1 of CaCO3 and CaCO3-MgCO3 respectively. Current and potencial pH were determined potentiometrically in a 1:2,5 water and 1:1 KCl relation respectively. Also cationic exchange capacity (CEC) with 1M NH4 acetate pH 7 as extractant and cationic exchange capacity not buffered (CICnb) with 0.2 M NH4Cl extractant at soil´s pH were determined. The four protocols used to evaluate the pH were highly associated. The four pH measures were able to discriminate liming rates although the current pH 1:2,5 presented the greatest differences. However, actual pH measurements were more volatile than the potential pH, at any dilution. The results indicate that 6-month anticipation liming may be sufficient for sensitive crops in soils with similar pH to those studied here. None of the determination methods for the cation exchange capacity and the saturation of the bases resulted in a size increase before liming or to discriminate the effect of each rate.Facultad de Ciencias Agrarias y Forestale

Antioxidant vitamin and mineral supplements for slowing the progression of age-related macular degeneration

Background: It has been proposed that antioxidants may prevent cellular damage in the retina by reacting with free radicals that are produced in the process of light absorption. Higher dietary levels of antioxidant vitamins and minerals may reduce the risk of progression of age-related macular degeneration (AMD). Objectives: The objective of this review was to assess the effects of antioxidant vitamin or mineral supplementation on the progression of AMD in people with AMD. Search methods: We searched CENTRAL (2017, Issue 2), MEDLINE Ovid (1946 to March 2017), Embase Ovid (1947 to March 2017), AMED (1985 to March 2017), OpenGrey (System for Information on Grey Literature in Europe, the ISRCTN registry (www.isrctn.com/editAdvancedSearch), ClinicalTrials.gov (www.clinicaltrials.gov) and the WHO International Clinical Trials Registry Platform (ICTRP) (www.who.int/ictrp/search/en). We did not use any date or language restrictions in the electronic searches for trials. We last searched the electronic databases on 29 March 2017. Selection criteria: We included randomised controlled trials (RCTs) that compared antioxidant vitamin or mineral supplementation (alone or in combination) to placebo or no intervention, in people with AMD. Data collection and analysis: Both review authors independently assessed risk of bias in the included studies and extracted data. One author entered data into RevMan 5; the other author checked the data entry. We graded the certainty of the evidence using GRADE. Main results: We included 19 studies conducted in USA, Europe, China, and Australia. We judged the trials that contributed data to the review to be at low or unclear risk of bias. Nine studies compared multivitamins with placebo (7 studies) or no treatment (2 studies) in people with early and moderate AMD. The duration of supplementation and follow-up ranged from nine months to six years; one trial followed up beyond two years. Most evidence came from the Age-Related Eye Disease Study (AREDS) in the USA. People taking antioxidant vitamins were less likely to progress to late AMD (odds ratio (OR) 0.72, 95% confidence interval (CI) 0.58 to 0.90; 2445 participants; 3 RCTs; moderate-certainty evidence). In people with very early signs of AMD, who are at low risk of progression, this would mean that there would be approximately 4 fewer cases of progression to late AMD for every 1000 people taking vitamins (1 fewer to 6 fewer cases). In people at high risk of progression (i.e. people with moderate AMD) this would correspond to approximately 8 fewer cases of progression for every 100 people taking vitamins (3 fewer to 13 fewer). In one study of 1206 people, there was a lower risk of progression for both neovascular AMD (OR 0.62, 95% CI 0.47 to 0.82; moderate-certainty evidence) and geographic atrophy (OR 0.75, 95% CI 0.51 to 1.10; moderate-certainty evidence) and a lower risk of losing 3 or more lines of visual acuity (OR 0.77, 95% CI 0.62 to 0.96; 1791 participants; moderate-certainty evidence). Low-certainty evidence from one study of 110 people suggested higher quality of life scores (National Eye Institute Visual Function Questionnaire) in treated compared with the non-treated people after 24 months (mean difference (MD) 12.30, 95% CI 4.24 to 20.36). Six studies compared lutein (with or without zeaxanthin) with placebo. The duration of supplementation and follow-up ranged from six months to five years. Most evidence came from the AREDS2 study in the USA. People taking lutein or zeaxanthin may have similar or slightly reduced risk of progression to late AMD (RR 0.94, 95% CI 0.87 to 1.01; 6891 eyes; low-certainty evidence), neovascular AMD (RR 0.92, 95% CI 0.84 to 1.02; 6891 eyes; low-certainty evidence), and geographic atrophy (RR 0.92, 95% CI 0.80 to 1.05; 6891 eyes; low-certainty evidence). A similar risk of progression to visual loss of 15 or more letters was seen in the lutein and control groups (RR 0.98, 95% CI 0.91 to 1.05; 6656 eyes; low-certainty evidence). Quality of life (measured with Visual Function Questionnaire) was similar between groups in one study of 108 participants (MD 1.48, 95% -5.53 to 8.49, moderate-certainty evidence). One study, conducted in Australia, compared vitamin E with placebo. This study randomised 1204 people to vitamin E or placebo, and followed up for four years. Participants were enrolled from the general population; 19% had AMD. The number of late AMD events was low (N = 7) and the estimate of effect was uncertain (RR 1.36, 95% CI 0.31 to 6.05, very low-certainty evidence). There were no data on neovascular AMD or geographic atrophy.There was no evidence of any effect of treatment on visual loss (RR 1.04, 95% CI 0.74 to 1.47, low-certainty evidence). There were no data on quality of life. Five studies compared zinc with placebo. The duration of supplementation and follow-up ranged from six months to seven years. People taking zinc supplements may be less likely to progress to late AMD (OR 0.83, 95% CI 0.70 to 0.98; 3790 participants; 3 RCTs; low-certainty evidence), neovascular AMD (OR 0.76, 95% CI 0.62 to 0.93; 2442 participants; 1 RCT; moderate-certainty evidence), geographic atrophy (OR 0.84, 95% CI 0.64 to 1.10; 2442 participants; 1 RCT; moderate-certainty evidence), or visual loss (OR 0.87, 95% CI 0.75 to 1.00; 3791 participants; 2 RCTs; moderate-certainty evidence). There were no data reported on quality of life. Very low-certainty evidence was available on adverse effects because the included studies were underpowered and adverse effects inconsistently reported. Authors' conclusions: People with AMD may experience some delay in progression of the disease with multivitamin antioxidant vitamin and mineral supplementation. This finding was largely drawn from one large trial, conducted in a relatively well-nourished American population. We do not know the generalisability of these findings to other populations. Although generally regarded as safe, vitamin supplements may have harmful effects. A systematic review of the evidence on harms of vitamin supplements is needed. Supplements containing lutein and zeaxanthin are heavily marketed for people with age-related macular degeneration but our review shows they may have little or no effect on the progression of AMD

Effects of phytogenic compounds on growth and nutritional physiology of Nile tilapia (Oreochromis niloticus)

With increasing world population, the demand for fish is growing thus there is a need to identify products with potential to increase the efficiency of fish production. Phytogenics are among the products being investigated as potential naturally derived growth promoters. The aim of this study was to identify phytogenic compounds and doses with growth-promoting effects in Nile tilapia and investigate relevant pathways underlying their growth promotion effects. The phytogenic compounds limonene, carvacrol and thymol, major constituents of essential oils from the plants citrus, oregano and thyme, respectively, were evaluated. Six Trials (Trials I, II, III, IV, V and VI) were carried out using diets supplemented with varying concentrations of the phytogenic compounds. In Trials I, II and III (Chapter 3), the effects of either limonene (Trial I), carvacrol (Trial II) or thymol (Trial III) on growth performance of Nile tilapia were investigated (objective 1) and performance parameters including final fish weight, daily growth coefficient, growth rate per metabolic body weight, percentage (%) weight gain, % survival, feed intake, feed conversion ratio and protein efficiency ratio were evaluated. Results from Trials I, II and III indicated that dietary supplementation of 400 and 500 ppm limonene and 750 ppm thymol had growth-promoting effects in Nile tilapia but the somatic growth was not associated with enhanced feed intake and feed utilisation efficiency. Trials IV and V (Chapter 4) investigated growth and nutritional physiology pathways in Nile tilapia regulated by individual phytogenic compounds (objective 2). This was accomplished by analysing the effects of limonene (Trial IV) and thymol (Trial V) supplemented diets on the expression of key genes participating in selected pathways of somatotropic axis-mediated growth, appetite regulation, nutrient digestion, absorption and transport, lipid metabolism, and antioxidant enzyme defence system. Limonene was supplemented in the diet at 0, 200, 400 and 600 ppm while thymol was supplemented at 0, 250 and 500 ppm. Trials IV and V found that growth-promoting effects of limonene (400 and 600 ppm) in Nile tilapia involved up-regulation of key genes within pathways including somatotropic axis-mediated growth, nutrient digestion, absorption and transport, lipid metabolism and antioxidant enzyme defence system. Dietary thymol at 250 and 500 ppm did not significantly enhance growth of Nile tilapia nor regulate the nutritional physiology pathways listed above. In Trial VI (Chapter 5), the effects of combined phytogenic compounds (limonene and thymol) on growth and nutritional physiology of Nile tilapia was tested (objective 3) to establish if the compounds had synergistic or additive effects on the growth of the fish as well as complementary effects on the selected nutritional physiology pathways. A candidate gene approach was also used for the selected pathways. Results from Trial VI showed that a diet supplemented with a combination of limonene (400 ppm) and thymol (500 ppm) has neither synergistic nor additive effects on the growth performance of Nile tilapia, with limonene mainly influencing the attained somatic growth. The analysed candidate genes involved in the pathways of nutrient digestion, absorption and transport, lipid metabolism, antioxidant enzymes and somatotropic axis growth also showed no synergistic or additive effects of a dietary combination of limonene and thymol in Nile tilapia. Overall, results from the study suggest approaches for developing functional diets for Nile tilapia using limonene and thymol growth promoters

Estratégias de controle paranematoide das galhas (Meloidogyne incognita) no cafeeiro.

O nematoideMeloidogyne incognitaconstituium dos principais fatores limitantes ao estabelecimento e a sustentabilidade das lavouras cafeeirasno estado de Rondônia.O controle químico (nematicida) tem sido inviável para o cafeeiro, uma vez que, seu uso torna-sefrequente em áreas infestadas, causando assim alto custo econômico e ambiental. Visando alternativasde controle que possa minimizar e ou substituir o uso de nematicidas químicos,objetivou-se nesse trabalho testar estratégias de controle contra M.incognitano cafeeiro. Este estudo testou três estratégiasde contole, sendo:Caracterização de plantasresistentes a M. incognitacomponentes dos genótiposdo programa de melhoramento genético da Embrapa Rondônia;Controle bilógico utilizando os micororganismos,Glomus macrocarpum(Fungos Micorrizicos Arbusculares-FMA) e cepas da rizobactéria Bacillus cereus; Controlecomextratosaquosos de plantas dos gêneros Mucunae Crotalaria. Os ensaiosforam conduzidos em casa de vegetaçãoem mudas de cafeeiro com seis meses, conduzidasindividualmente em vasos de 8 L,inoculadas com 5.000 ovos.Os parâmetros avaliados foram: número de galhas por grama de raiz(NG), número de ovos e juvenis(J2)por planta(NO)e fator de reprodução(FR). Os resultados obtidos foram: Para Controle genético,seis genótipos(BRS3210, C12, BRS2299, BRS2314, BRS3137 e BRS1216) idetificados como resistentes. Para controle biológico os tratamentos: micorriza isolado, rizobactérias isolado (estirpesRz216, Rz216 e Rz48), e interação (rizobactérias+ micorriza), na presença de nematoide, mostratam eficiência de controle similres ao tratamento nematicida.Para os extratos aquosos, observou-se que: Crotalaria juncearamo seco (CJRS), C. ochroleucafolhaseca (COFLS), C. espectabilisfolha seca (CEFlS), Mucuna preta ramoseco (MPRS), C. junceafruto fresco (CJFrF), C. espectabilisfruto fresco (CEFrF), Mucuna preta folha seca (MPFlS), C. ochroleucaramo fresco (CORF) e C. espectabilisfruto seco (CEFrS) expressaram resultado similar ao nematicida. Observou-se que as estratégias avaliadas se mostraram eficientes para o controle de M. incognitaem cafeeiro. Estudos posteriores são necessários para identificação dos mecanismos de ação e constituintes químicosativos, viabilizando, assim, novas alternativas de controle para o nematóide das galhas no cafeeiro na região Amazônica.Tese (Doutorado em Biodiversidade e Conservação) - Fundação Universidade Federal de Rondônia, Rondônia. Orientador: Cleberson de Freitas Fernandes, Co-orientador: José Roberto Vieira Junior